Journal: bioRxiv

Article Title: Red yeast rice-derived MKA ameliorates cardiac hypertrophy in hypertensive rats by inhibiting ERK1/2/c-Fos pathway

doi: 10.64898/2026.03.10.710945

Figure Lengend Snippet: Representative images of EDU and TUNEL staining ( A ), EDU positive percent ( B ) and apoptosis percent ( C ) in H9c2 cardiomyocytes treated with different concentrations of MKA (1, 5, 10 μM) and 200 nM Ang II for 24 hours. D-H. Representative Western blots and relative protein expression for β-MHC, p-ERK, t-ERK, c-Fos, and Caspase-3. Data are presented as mean ± SE, comparisons among multiple groups were conducted using one-way analysis of variance (ANOVA) followed by Tukey’s multiple comparison test (n = 4 per group; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001). LM: 1 μM MKA; MM: 5 μM MKA; HM: 10 μM MKA;

Article Snippet: Cells were fixed with 4% paraformaldehyde, permeabilized with 0.3% Triton X-100/PBS (VICMED) for 5 minutes, and processed using the One-step TUNEL In Situ Apoptosis Kit (Red, Elab Fluor® 594) (Elabscience Co., Ltd., Wuhan, E-CK-A322) according to the manufacturer’s instructions.

Techniques: TUNEL Assay, Staining, Western Blot, Expressing, Comparison

Journal: Frontiers in Immunology

Article Title: Neuroinflammatory and transcriptional dynamics during SARS-CoV-2 infection in KRT18-hACE2 mouse brain

doi: 10.3389/fimmu.2026.1716597

Figure Lengend Snippet: Neuropathological changes and inflammatory mediators in K18-hACE mouse brain following SARS-CoV-2 infection. (A, B) Immunofluorescence staining of brain tissue from uninfected and SARS-CoV-2 infected mice showing colocalization of DAPI (blue, nuclear stain) with (A) Iba-1 (green) and (B) CD68 (red). Lower panels show single staining for Iba-1 or CD68. Scale bar: 50 µm. (C) TUNEL assay of brain tissue from uninfected and SARS-CoV-2 infected mice. (D) Quantification of panel C showing the percentage of TUNEL+ cells in each group. Student t-test was performed to determine the significance. **p < 0.01, ***p < 0.001).

Article Snippet: DNA fragmentation was detected on rehydrated paraffin-embedded sections using a one-step TUNEL in situ apoptosis detection kit (E-CK-A325; Elabscience), according to the manufacturer’s instructions.

Techniques: Infection, Immunofluorescence, Staining, TUNEL Assay

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: lncRNA NEAT1 promotes hypoxia‐induced inflammation and fibrosis of alveolar epithelial cells via targeting miR ‐29a/ NFATc3 axis

doi: 10.1002/kjm2.12535

Figure Lengend Snippet: Knockdown of NEAT1 alleviated hypoxia‐stimulated inflammatory responses, apoptosis and fibrosis in pulmonary alveolar epithelial cells. HPAEpic and A549 cells were transfected with NEAT1 shRNA (sh‐NEAT1) and negative control shRNA (sh‐NC) and then exposed to hypoxic stimulation. (A) qRT–PCR analysis of NEAT1 levels. (B) Cell apoptosis was analyzed by a TUNEL assay. (C) The levels of inflammatory factors, including TNF‐α, IL‐1β, and IL‐6, in HPAEpic and A549 cells were analyzed by ELISAs. (D) Western blot analysis of the expression of the fibrosis‐associated proteins, including α‐SMA, collagen I and collagen III. Data represent the mean ± SD ( n = 3 independent experiments). * p < 0.05; ** p < 0.01; *** p < 0.001. Hx, Hypoxia; Nx, Normoxia

Article Snippet: Cell apoptosis was monitored by TUNEL Apoptosis Assay Kits (Beyotime Biotech).

Techniques: Knockdown, Transfection, shRNA, Negative Control, Quantitative RT-PCR, TUNEL Assay, Western Blot, Expressing

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: lncRNA NEAT1 promotes hypoxia‐induced inflammation and fibrosis of alveolar epithelial cells via targeting miR ‐29a/ NFATc3 axis

doi: 10.1002/kjm2.12535

Figure Lengend Snippet: miR‐29a/NFATc3 axis was participated in the regulation of NEAT1 in hypoxia‐induced inflammation and fibrosis. HPAEpic and A549 cells were transfected with sh‐NEAT1 + miR‐29a inhibitor or sh‐NEAT1 + pcDNA‐NFATc3 and further exposed to hypoxic stimulation. (A,B) qRT–PCR analyzed the expression of miR‐29a and NFATc3 in HPAEpic and A549 cells. (C) TUNEL assays detected the cell apoptosis. (D) The levels of inflammatory factors, including TNF‐α, IL‐1β, and IL‐6, in HPAEpic and A549 cells were detected by ELISAs. (E) Western blot analysis of the expression of the fibrosis‐associated proteins, including α‐SMA, collagen I and collagen III. Data represent the mean ± SD ( n = 3 independent experiments). * p < 0.05; ** p < 0.01; *** p < 0.001. Hx, Hypoxia; Nx, Normoxia

Article Snippet: Cell apoptosis was monitored by TUNEL Apoptosis Assay Kits (Beyotime Biotech).

Techniques: Transfection, Quantitative RT-PCR, Expressing, TUNEL Assay, Western Blot

Journal: Journal of Nanobiotechnology

Article Title: Targeting delivery of miR-146a via IMTP modified milk exosomes exerted cardioprotective effects by inhibiting NF-κB signaling pathway after myocardial ischemia-reperfusion injury

doi: 10.1186/s12951-024-02631-0

Figure Lengend Snippet: MEs-miR-146a protects H9c2 cells from OGD/R induced damage. ( A ) After 24 h of incubation, PKH26 labeled exosomes could be uptaken up by H9c2. Scale bar = 20 μm. ( B ) MEs-miR-146a decreased OGD/R induced cell apoptosis of H9c2 according to TUNEL assay. Scale bar = 50 μm. * P < 0.05, ** P < 0.01 versus the control group; & P < 0.05, && P < 0.01 versus the OGD/R group; ▲ P < 0.05 versus the OGD/R-MEs group. n = 6

Article Snippet: The one-step TUNEL Apoptosis Detection Kit (C1088, Green Fluorescence, Beyotime) was used to incubate for 60 min at RT, following by nuclear labeling with DAPI (H-1200-10, Vector Laboratories, Burlingame, CA, USA), both in the dark.

Techniques: Incubation, Labeling, TUNEL Assay, Control

Journal: Journal of Nanobiotechnology

Article Title: Targeting delivery of miR-146a via IMTP modified milk exosomes exerted cardioprotective effects by inhibiting NF-κB signaling pathway after myocardial ischemia-reperfusion injury

doi: 10.1186/s12951-024-02631-0

Figure Lengend Snippet: MEs-miR-146a protects NRCM cells from OGD/R induced damage. ( A ) Representative fluorescence images indicated that PKH26 labeled exosomes were taken up by NRCM cells after 24 h of incubation. Scale bar = 20 μm. ( B ) MEs-miR-146a decreased OGD/R induced cell apoptosis of NRCM according to TUNEL assay. Scale bar = 50 μm. ** P < 0.01 versus the control group; & P < 0.05, && P < 0.01 versus the OGD/R group; ▲ P < 0.05 versus the OGD/R-MEs group. n = 6

Article Snippet: The one-step TUNEL Apoptosis Detection Kit (C1088, Green Fluorescence, Beyotime) was used to incubate for 60 min at RT, following by nuclear labeling with DAPI (H-1200-10, Vector Laboratories, Burlingame, CA, USA), both in the dark.

Techniques: Fluorescence, Labeling, Incubation, TUNEL Assay, Control

Journal: Journal of Nanobiotechnology

Article Title: Targeting delivery of miR-146a via IMTP modified milk exosomes exerted cardioprotective effects by inhibiting NF-κB signaling pathway after myocardial ischemia-reperfusion injury

doi: 10.1186/s12951-024-02631-0

Figure Lengend Snippet: MEs-miR-146a exhibited better therapeutic efficacy in decreasing apoptotic cells and limiting inflammation than MEs. ( A - B ) Representative TUNEL staining images and quantification of apoptotic radio. ** P < 0.01 versus the Sham group; & P < 0.05 and && P < 0.01 versus the MIRI group. ( C - F ) The expression of inflammatory factors of rat serum were detected by ELISA. * P < 0.05, ** P < 0.01 versus the Sham group; & P < 0.05, && P < 0.01 versus the MIRI group; ▲ P < 0.05, ▲▲ P < 0.01 versus the MIRI-MEs-miR-146a group. n = 5

Article Snippet: The one-step TUNEL Apoptosis Detection Kit (C1088, Green Fluorescence, Beyotime) was used to incubate for 60 min at RT, following by nuclear labeling with DAPI (H-1200-10, Vector Laboratories, Burlingame, CA, USA), both in the dark.

Techniques: Drug discovery, TUNEL Assay, Staining, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Journal of Nanobiotechnology

Article Title: Targeting delivery of miR-146a via IMTP modified milk exosomes exerted cardioprotective effects by inhibiting NF-κB signaling pathway after myocardial ischemia-reperfusion injury

doi: 10.1186/s12951-024-02631-0

Figure Lengend Snippet: Tail vein injection of IMTP-MEs-miR-146a significantly ameliorated myocardium apoptosis and attenuated inflammation at the early stage of MIRI. ( A ) Representative H&E images of heart tissue from each group. Scale bar = 50 μm. ( B - C ) Representative images of TUNEL staining 24 h after MIRI and quantitative analysis of apoptotic radio. * P < 0.05 and ** P < 0.01 versus the MIRI group; & P < 0.05 versus the MIRI-MEs-miR-146a group. n = 8. ( D - G ) The expression of inflammatory factors of rat serum were detected by ELISA. ∆∆ P < 0.01 versus the MIRI group; ▲▲ P < 0.01 versus the MIRI-MEs-miR-146a group. n = 8

Article Snippet: The one-step TUNEL Apoptosis Detection Kit (C1088, Green Fluorescence, Beyotime) was used to incubate for 60 min at RT, following by nuclear labeling with DAPI (H-1200-10, Vector Laboratories, Burlingame, CA, USA), both in the dark.

Techniques: Injection, TUNEL Assay, Staining, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Acta pharmaceutica Sinica. B

Article Title: A cyclodextrin-based nanoformulation achieves co-delivery of ginsenoside Rg3 and quercetin for chemo-immunotherapy in colorectal cancer.

doi: 10.1016/j.apsb.2021.06.005

Figure Lengend Snippet: Figure 2 Ginsenoside Rg3 induced immunogenic cell death in CRC cells. (A) IC50 of Rg3 for CT26 and HCT116 cells at 24 h. Data are presented as mean SD (n Z 3). (B) Apoptosis (%) in CT26 and HCT116 cells following treatment of Rg3 at 6, 12 and 24 h. Data are presented as mean SD (n Z 3). *P < 0.05 and **P < 0.01 relative to DMSO. (C) The activity of UPR signaling pathways following treatment of Rg3 ([c] Z 30 mmol/L) at 6, 12 and 24 h. The quantification was demonstrated in Fig. S1. (D) The characterization of ICD in CRC cells following treatment of Rg3 ([c] Z 30 mmol/L), including CRT exposure (6 h), ATP secretion (12 h) and HMGB1 release (12 h). Data are presented as mean SD (n Z 3). *P < 0.05 relative to DMSO; scale bar Z 20 mm. (E) The expression of CD11c and CD86 in DCs stimulated with the supernatant from Rg3 ([c] Z 30 mmol/L) -treated cells (24 h). Data are presented as mean SD (n Z 3). **P < 0.01 relative to DMSO. (F) The in vivo vaccination assay using BALB/C and nude mice. Data are presented as mean SD (n Z 4). ***P < 0.001.

Article Snippet: DNA fragments were detected using the TransDetect Fluorescein TUNEL Cell Apoptosis Kit (TransGen Biotech), and nuclei were stained with DAPI (Beyotime Biotech), for confocal microscopic analysis (Olympus); 2) Measurement of immune cells.

Techniques: Activity Assay, Protein-Protein interactions, Expressing, In Vivo

Journal: Acta pharmaceutica Sinica. B

Article Title: A cyclodextrin-based nanoformulation achieves co-delivery of ginsenoside Rg3 and quercetin for chemo-immunotherapy in colorectal cancer.

doi: 10.1016/j.apsb.2021.06.005

Figure Lengend Snippet: Figure 3 Quercetin caused reactive oxygen species in CRC cells. (A) IC50 of QTN for CT26 and HCT116 cells at 24 h. Data are presented as mean SD (n Z 3). (B) Apoptosis (%) in CT26 and HCT116 cells following treatment of QTN ([c] Z 80 mmol/L) at 24 h. Data are presented as mean SD (n Z 3). **P < 0.01 relative to DMSO. (C) The activity of Bcl-2/BAX/caspase 9/caspase 3 signaling pathways following treatment of QTN ([c] Z 80 mmol/L) at 6, 12 and 24 h. The quantification was demonstrated in Fig. S2. (D) The ROS level in CT26 and HCT116 cells following treatment of QTN ([c] Z 80 mmol/L) at 6, 12 and 24 h. Data are presented as mean SD (n Z 3). **P < 0.01 and ***P < 0.001 relative to DMSO. (E) Cell viability (%) of CT26 and HCT116 cells with or without NAC prior to treatment of QTN ([c] Z 80 mmol/L) (24 h). Data are presented as mean SD (n Z 3). *P < 0.05 and **P < 0.01 relative to untreated control. (F) Apoptosis (%) in CT26 and HCT116 cells with or without NAC prior to treatment of QTN ([c] Z 80 mmol/L) (24 h). Data are presented as mean SD (n Z 3). *P < 0.05 and **P < 0.01 relative to untreated control.

Article Snippet: DNA fragments were detected using the TransDetect Fluorescein TUNEL Cell Apoptosis Kit (TransGen Biotech), and nuclei were stained with DAPI (Beyotime Biotech), for confocal microscopic analysis (Olympus); 2) Measurement of immune cells.

Techniques: Activity Assay, Protein-Protein interactions, Control

Journal: Acta pharmaceutica Sinica. B

Article Title: A cyclodextrin-based nanoformulation achieves co-delivery of ginsenoside Rg3 and quercetin for chemo-immunotherapy in colorectal cancer.

doi: 10.1016/j.apsb.2021.06.005

Figure Lengend Snippet: Figure 4 Synergistic effects of Rg3 and QTN in CT26 cells. (A) IC50 of drug combination at 24 h. Data are presented as mean SD (n Z 3). CI values at IC50 were shown in Fig. S3. (B) Apoptosis (%) caused by drug combination at 24 h. Data are presented as mean SD (n Z 3). *P < 0.05 and **P < 0.01 relative to DMSO. (C) The CRT exposure with or without NAC before treatment of drug combination (6 h). Data are presented as mean SD (n Z 3). *P < 0.05 and **P < 0.01 relative to DMSO; scale bar Z 20 mm. (D) The ATP secretion with or without NAC before treatment of drug combination at 12 h. Data are presented as mean SD (n Z 3).*P < 0.05 and **P < 0.01, between NAC and No NAC. (E) The HMGB1 release with or without NAC before treatment of drug combination at 12 h. Data are presented as mean SD (n Z 3). *P < 0.05 and **P < 0.01, between NAC and No NAC. (F) The expression of CD11c and CD86 in DCs stimulated (24 h) by the supernatant from Rg3- treated cells with or without pretreatment of NAC. Data are presented as mean SD (n Z 3). *P < 0.05, **P < 0.01 and ***P < 0.001, relative to DMSO.

Article Snippet: DNA fragments were detected using the TransDetect Fluorescein TUNEL Cell Apoptosis Kit (TransGen Biotech), and nuclei were stained with DAPI (Beyotime Biotech), for confocal microscopic analysis (Olympus); 2) Measurement of immune cells.

Techniques: Expressing

Journal: Acta pharmaceutica Sinica. B

Article Title: A cyclodextrin-based nanoformulation achieves co-delivery of ginsenoside Rg3 and quercetin for chemo-immunotherapy in colorectal cancer.

doi: 10.1016/j.apsb.2021.06.005

Figure Lengend Snippet: Figure 8 Combination therapy of targeted co-formulation and Anti-PD-L1 for CRC. (A) Treatment schedule and IVIS images. (B) The CRC progression over a 35-day period. Data are presented as mean SD (n Z 5). *P < 0.05 and **P < 0.01; NS, no significance. (C) Animal survival (median survival: PBS ~38 days, Anti-PD-L1 ~40 days, targeted co-formulation ~62 days, and combination z 96 days). Data are presented as mean SD (n Z 5). **P < 0.01 and ***P < 0.001. (D) Immunofluorescent staining assay (green Z DNA fragments and blue Z nuclei) on Day 20 to assess apoptosis in the tumor (scale bar Z 50 mm). Data are presented as mean SD (n Z 3). *P < 0.05 and **P < 0.01, relative to PBS. (E) Level of immune cells in the tumor on Day 20 was analyzed using flow cytometry (BD). Data are presented as mean SD (n Z 4). *P < 0.05 and **P < 0.01; NS, no significance. (F) The mRNA expression of cytokines and chemokines in the tumor on Day 20 was analyzed using real time RT-PCR. Data are presented as mean SD (n Z 4). *P < 0.05 and **P < 0.01; NS, no significance. (G) Orthotopic CRC mice treated with targeted co-formulation following the removal of CD4 þ or CD8þ T cells. Data are presented as mean SD (n Z 4). *P < 0.05 and **P < 0.01; NS, no significance.

Article Snippet: DNA fragments were detected using the TransDetect Fluorescein TUNEL Cell Apoptosis Kit (TransGen Biotech), and nuclei were stained with DAPI (Beyotime Biotech), for confocal microscopic analysis (Olympus); 2) Measurement of immune cells.

Techniques: Formulation, Staining, Cytometry, Expressing, Quantitative RT-PCR

Journal: Drug Design, Development and Therapy

Article Title: Oxysophocarpine Inhibits Apoptosis of Lung Epithelial Cells to Alleviate Acute Lung Injury via KIT/PI3K Signaling Pathway

doi: 10.2147/DDDT.S544479

Figure Lengend Snippet: OSC attenuates histopathological changes in lungs of mice with acute lung injury. ( A ) HE staining detection of mouse air lung tissue sections. Magnification ×20. The red arrow indicates alveolar collapse and diffuse inflammatory cell infiltration. ( B ) Lung wet to dry weight ratio. ( C ) Total number of cells in BALF. ( D ) APC-FITC double staining to detect neutrophil accumulation in BALF. ( E ) Total number of neutrophil aggregates in BALF. ( F ) Levels of TNF-α in mouse BALF were determined by ELISA. ( G ) ELISA to determine the level of IL-1β in mouse BALF. ( H ) ELISA to determine the level of IL-6 in mouse BALF. Values are expressed as mean ± SD (n = 5). Compared with the Normal group, *p < 0.05, **p < 0.01; compared with the LPS model group, # p < 0.05, ## p < 0.01. OSC exhibited inhibition of apoptosis in an ALI mouse model.

Article Snippet: According to the instructions provided in the TUNEL cell apoptosis assay kit (SF594, 20231008, Solarbio, Beijing, China), the paraffin sections were deparaffinized twice in xylene and then subjected to a gradient treatment with ethanol-water solutions.

Techniques: Staining, Double Staining, Enzyme-linked Immunosorbent Assay, Inhibition

Journal: Drug Design, Development and Therapy

Article Title: Oxysophocarpine Inhibits Apoptosis of Lung Epithelial Cells to Alleviate Acute Lung Injury via KIT/PI3K Signaling Pathway

doi: 10.2147/DDDT.S544479

Figure Lengend Snippet: OSC exhibits inhibition of apoptosis in an ALI mouse model. ( A ) TUNEL staining to observe apoptosis of lung tissue cells. Magnification × 20. ( B ) Fluorescence intensity was analysed using Image J software. Values are expressed as mean ± SD (n = 5). **p < 0.01 compared with Normal group; ## p < 0.01 compared with LPS model group.

Article Snippet: According to the instructions provided in the TUNEL cell apoptosis assay kit (SF594, 20231008, Solarbio, Beijing, China), the paraffin sections were deparaffinized twice in xylene and then subjected to a gradient treatment with ethanol-water solutions.

Techniques: Inhibition, TUNEL Assay, Staining, Fluorescence, Software

Journal: Drug Design, Development and Therapy

Article Title: Oxysophocarpine Inhibits Apoptosis of Lung Epithelial Cells to Alleviate Acute Lung Injury via KIT/PI3K Signaling Pathway

doi: 10.2147/DDDT.S544479

Figure Lengend Snippet: OSC Alleviates LPS-Induced Apoptosis of BEAS-2B Cells. ( A ) MTT assay to determine the effect of OSC on cell viability (n=5). ( B ) MTT assay to determine the effect of LPS on cell viability (n=5). ( C ) Annexin V/PI double staining to detect apoptosis of pulmonary epithelial cells. Values are expressed as mean±SD (n=3). *p < 0.05 compared with the Normal group; ## p < 0.01 compared with the LPS model group.

Article Snippet: According to the instructions provided in the TUNEL cell apoptosis assay kit (SF594, 20231008, Solarbio, Beijing, China), the paraffin sections were deparaffinized twice in xylene and then subjected to a gradient treatment with ethanol-water solutions.

Techniques: MTT Assay, Double Staining

Journal: Drug Design, Development and Therapy

Article Title: Oxysophocarpine Inhibits Apoptosis of Lung Epithelial Cells to Alleviate Acute Lung Injury via KIT/PI3K Signaling Pathway

doi: 10.2147/DDDT.S544479

Figure Lengend Snippet: Network pharmacological analysis and molecular docking predicted the potential mechanism of OSC in alleviating epithelial cell apoptosis to treat acute lung injury. ( A ) Venn diagram of common targets between acute lung injury and pulmonary epithelial cell apoptosis. ( B ) KEGG pathway enrichment. ( C ) PPI protein interaction core target analysis. ( D ) Mechanism diagram of OSC alleviating epithelial cell apoptosis and reducing acute lung injury. ( E ) Molecular docking of OSC with KIT, binding energy of −6.32 kJ/mol. ( F ) Molecular docking of OSC with PIK3CA, binding energy of −7.84 kJ/mol. ( G ) Molecular docking of OSC with Bcl-2, binding energy of −7.92 kJ/mol.

Article Snippet: According to the instructions provided in the TUNEL cell apoptosis assay kit (SF594, 20231008, Solarbio, Beijing, China), the paraffin sections were deparaffinized twice in xylene and then subjected to a gradient treatment with ethanol-water solutions.

Techniques: Binding Assay

Journal: Ecotoxicology and environmental safety

Article Title: Autophagy promotes GSDME-mediated pyroptosis via intrinsic and extrinsic apoptotic pathways in cobalt chloride-induced hypoxia reoxygenation-acute kidney injury.

doi: 10.1016/j.ecoenv.2022.113881

Figure Lengend Snippet: Fig. 3. Apoptosis was induced in the PTECs in the CoCl2-induced HRI-AKI models. (A) Representative of the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assays for the apoptotic cells in the kidney samples (Scale bar: 50 µm). The nuclei of the TUNEL-positive cells were stained green. (B) The TUNEL results in HK-2 cells. Scale bar: 100 µm. (C) The flow cytometry assays detecting the apoptotic cells in HK-2 cells treated with different concentrations of CoCl2 for 24 h and then reoxygenated for 6 h. (D) The quantitative ratios of the apoptotic cells and necrotic cells in 3 C. (E) The expression levels of caspase 8 (CASP 8), cleaved caspase 8 (c-CASP 8), CASP 9, c-CASP 9, CASP 3 and c-CASP 3 detected with western blotting assays. The protein samples were prepared from the mouse kidneys after IRI treatment. (F) The quantitative results of the relative expression levels of c-CASP 8, c-CASP 9 and c-CASP 3 in E. (G) The western blotting assays detecting the apoptosis-related proteins in HK-2 cells treated with different concentrations of CoCl2 for 24 h and then reoxygenated for 6 h. (H) The quantitative results of the relative expression levels of c-CASP 8, c-CASP 9 and c-CASP 3 in Fig. G. (I) The western blotting assays detecting the apoptosis-related proteins in HK-2 cells treated with 500 μM CoCl2 for 24 h and then reoxygenated for different reoxygenation times. (J) The quantitative results of the relative expression levels of c-CASP 8, c-CASP 9 and c-CASP 3 in I. The western blotting assays were repeated at least three times. *p < 0.05, **p < 0.01, and *** p < 0.001 compared with the Ctrl group, # p < 0.05, ## p < 0.01, and ### p < 0.001 compared with the hypoxia group with reoxygenation time of 0 h.

Article Snippet: To detect whether apoptosis occurred in HRI-AKI, the TUNEL apoptosis assay kit (Elabscience Biotechnology Co., Ltd, Wuhan, China) was used to detect DNA fragmentation according to the instruction.

Techniques: TUNEL Assay, Staining, Flow Cytometry, Expressing, Western Blot

Journal: Ecotoxicology and environmental safety

Article Title: Autophagy promotes GSDME-mediated pyroptosis via intrinsic and extrinsic apoptotic pathways in cobalt chloride-induced hypoxia reoxygenation-acute kidney injury.

doi: 10.1016/j.ecoenv.2022.113881

Figure Lengend Snippet: Fig. 4. Inhibition of apoptosis decreased GSDME-mediated pyroptosis in CoCl2- induced HRI-AKI models. (A) The effects of CASP 3 inhibitor, Z-DEVD-FMK, on the cell viabilities, the relative levels of ATP or the relative release of LDH induced in the CoCl2- induced HRI cell model. (B) The effects of CASP 3 inhibitor, Z-DEVD-FMK, on GSDME- mediated pyroptosis in HRI cell model. (C) The quantitative results of the relative expression levels of c-CASP 3 and GSDME-NT in B. (D) The effects of CASP 9 inhibitor, Z- LEHD-FMK TFA, on the cell viabilities, the relative levels of ATP or the relative release of LDH in the HRI cell model. (E) The effects of CASP 9 inhibitor, Z-LEHD-FMK TFA, on the protein levels of CASP 9, c-CASP 9, CASP 3, c-CASP 3, GSDME-FL, and GSDME-NT in the HRI cell model. (F) The quantitative re sults of the relative expression levels of c- CASP 9 and GSDME-NT in E. (G) The effects of CASP 8 inhibitor, Z-IETD-FMK, on the cell viabilities, the relative levels of ATP or the relative release of LDH in the HRI cell model. (H) The effects of CASP 8 inhibitor, Z-IETD- FMK, on the protein levels of CASP 8, c-CASP 8, CASP 9, c-CASP 9, CASP 3, c-CASP 3, GSDME-FL and GSDME-NT in HRI cell model. (I) The quantitative results of the relative expression levels of c-CASP 8 and GSDME-NT in H. *p < 0.05, **p < 0.01, and ***p < 0.001 compared with the corre sponding Ctrl group without HRI treatment. # p < 0.05, ## p < 0.01, and ### p < 0.001 compared with the Ctrl group with HRI treatment.

Article Snippet: To detect whether apoptosis occurred in HRI-AKI, the TUNEL apoptosis assay kit (Elabscience Biotechnology Co., Ltd, Wuhan, China) was used to detect DNA fragmentation according to the instruction.

Techniques: Inhibition, Expressing

Journal: Ecotoxicology and environmental safety

Article Title: Autophagy promotes GSDME-mediated pyroptosis via intrinsic and extrinsic apoptotic pathways in cobalt chloride-induced hypoxia reoxygenation-acute kidney injury.

doi: 10.1016/j.ecoenv.2022.113881

Figure Lengend Snippet: Fig. 6. The effects of inhibiting autophagy on pyroptosis and apoptosis in the HRI-AKI models. (A) The mRNA level of atg5 in HK-2 cells after lentivirus transfection to knock down Atg5. (B) The mRNA level of fip200 in HK-2 cells after lentivirus transfection to knock down FIP200. (C) The protein levels of Atg5 and FIP200 after lentivirus transfection to knock down Atg5 or FIP200. (D) The protein levels of LC3B-I and LC3B-II with HRI treatment after lentivirus transfection to knock down Atg5 or FIP200. (E) The densitometric analysis of LC3B-II protein levels in D. (F) The levels of related apoptotic and pyroptotic proteins in HK-2 cells after lentivirus transfection to knock down Atg5 or FIP200. (G) The densitometric analysis of c-CASP 8, c-CASP 9, c-CASP 3 and GSDME-NT in Fig. 6F. The data shown were repeated at least three times. *p < 0.05, **p < 0.01, and ***p < 0.001 compared with the corresponding Ctrl group without HRI treatment. ### p < 0.001 compared with the sh-ctrl group with HRI treatment.

Article Snippet: To detect whether apoptosis occurred in HRI-AKI, the TUNEL apoptosis assay kit (Elabscience Biotechnology Co., Ltd, Wuhan, China) was used to detect DNA fragmentation according to the instruction.

Techniques: Transfection, Knockdown